The synthesis of inhibitors for processing proteinases and their action on the Kex2 proteinase of yeast

Herbert Angliker; Peter Wikstrom; Elliott Shaw; Charles Brenner; Robert S Fuller

doi:10.1042/bj2930075

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The synthesis of inhibitors for processing proteinases and their action on the Kex2 proteinase of yeast

Journal article

Open access

Peer reviewed

The synthesis of inhibitors for processing proteinases and their action on the Kex2 proteinase of yeast

Herbert Angliker, Peter Wikstrom, Elliott Shaw, Charles Brenner and Robert S Fuller

The Biochemical journal, Vol.293 ( Pt 1)(1), pp.75-81

07/01/1993

DOI: 10.1042/bj2930075

PMID: 8328974

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https://www.ncbi.nlm.nih.gov/pmc/articles/1134322View

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Abstract

Peptidyl chloromethane and sulphonium salts containing multiple Arg and Lys residues were synthesized as potential inhibitors of prohormone and pro-protein processing proteinases. The potencies of these compounds were assayed by measuring the kinetics of inactivation of the yeast Kex2 proteinase, the prototype of a growing family of eukaryotic precursor processing proteinases. The most potent inhibitor, Pro-Nvl-Tyr-Lys-Arg-chloromethane, was based on cleavage sites in the natural Kex2 substrate pro-alpha-factor. This inhibitor exhibited a Ki of 3.7 nM and a second-order inactivation rate constant (k2/Ki) of 1.3 x 10(7) M-1.s-1 comparable with the value of kcat./Km obtained with Kex2 for the corresponding peptidyl methylcoumarinylamide substrate. The enzyme exhibited sensitivity to the other peptidyl chloromethanes over a range of concentrations, depending on peptide sequence and alpha-amino decanoylation, but was completely resistant to peptidyl sulphonium salts. Kinetics of inactivation by these new inhibitors of a set of 'control' proteinases, including members of both the trypsin and subtilisin families, underscored the apparent specificity of the compounds most active against Kex2 proteinase.

Amino Acid Sequence

Protease Inhibitors - pharmacology

Amino Acid Chloromethyl Ketones - chemical synthesis

Amino Acid Chloromethyl Ketones - pharmacology

Saccharomyces cerevisiae Proteins

Molecular Sequence Data

Protease Inhibitors - chemical synthesis

Kinetics

Proprotein Convertases

Yeasts - enzymology

Binding Sites

Subtilisins - antagonists & inhibitors

Details

Title: Subtitle: The synthesis of inhibitors for processing proteinases and their action on the Kex2 proteinase of yeast
Creators: Herbert Angliker - Friedrich Miescher-Institut, Basel, Switzerland
Peter Wikstrom
Elliott Shaw
Charles Brenner
Robert S Fuller
Resource Type: Journal article
Publication Details: The Biochemical journal, Vol.293 ( Pt 1)(1), pp.75-81
DOI: 10.1042/bj2930075
PMID: 8328974
NLM abbreviation: Biochem J
ISSN: 0264-6021
eISSN: 1470-8728
Publisher: England
Grant note: GM 39697 / NIGMS NIH HHS NRSA 5T32 CA 09302 / NCI NIH HHS
Language: English
Date published: 07/01/1993
Academic Unit: Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9983788596702771

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